Past studies in our laboratory have shown that physiological and morphological changes (activation) of Ascaris spermatozoa can be induced by subjecting them to homogenates of a male accessory sex gland called the glandular vas deferens. Objectives of the proposed study are to: (1) determine more precisely the morphological and chemical alterations which occur at the membrane level during the activation process, (2) determine the mechanism of activation and motility in the altered spermatozoa and, (3) develop an in vitro procedure utilizing Ascaris gametes. To determine the nature, density and distribution of carbohydrate rich residues present on the surface of both unactivated and activated cells, unlabeled Concanavalin A will be used as a marker. The position of the unlabeled Con-A on the membrane will be demonstrated using rabbit anti-Con A antisera, ferritin goat anti-rabbit IgG, and fluorescein labeled goat anti-rabbit IgG. To determine if the movement of certain protein particles from cytoplasmic membranous elements (following the latter's fusion with the plasma membrane) is responsible for newly acquired antigenic determinants on the surface of freshly activated spermatozoa, the cytoplasmic components will be isolated, and injected into rabbits. The recovered immunoglobulins will then be employed to determine if the membranous elements and the surface membrane of the activated sperm share common antigens. To determine if the motility of newly activated sperm depends upon the recruitment of microfilament precursors, fluorescein labeled HMM will be used to follow the distribution patterns of the microfilament assemblies following treatment of the sperm with glandular homogenates. Attempts to develop an in vitro fertilization procedure will be directed to placing sperm (both artificially activated as well as those recovered from the female uterus) and primary oocytes together in Media 199 under various gas phases and conditions which will most likely permit gamete membrane fusion.